LOCUS       pZIP-hCMV-ZSGree        7855 bp ds-DNA     circular SYN 19-DEC-2017
DEFINITION  synthetic circular DNA.
ACCESSION   . (modified)
VERSION     .
KEYWORDS    pZIP-hCMV-ZSGreen-Puro (V027).
SOURCE      null
  ORGANISM  
            .
REFERENCE   1  (bases 1 to 8373)
  AUTHORS   Transomic
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 16, 2016 from SnapGene 3.0.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     LTR             5356..5589
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (Delta-U3)"
     enhancer        2235..2538
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     gene            <3592..4188
                     /label="Puro"
     primer_bind     3496..3545
                     /label="3549p2apuro_F"
     misc_feature    complement(4170..>4205)
                     /note="Geneious type: primer_bind_reverse"
                     /label="p2apuro_R"
     misc_feature    3526..3591
                     /note="Geneious type: propeptide"
                     /label="P2A"
     rep_origin      complement(6103..6688)
                     /direction="LEFT"
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori rep origin"
     source          1..7855
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /label="source recombinant plasmid"
     CDS             complement(6859..7719)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="bla CDS"
     CDS             2833..>3525
                     /codon_start=1
                     /note="ZsGreen"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI
                     NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG
                     AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI
                     LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH
                     LTEHAIASGSALP"
                     /label="ZsGreen"
     promoter        5685..6014
                     /note="SV40"
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        2539..2742
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /label="CMV promoter"
     promoter        complement(7720..7824)
                     /gene="bla"
                     /note="AmpR promoter"
                     /label="bla promoter"
     misc_feature    23..667
                     /note="CMV-LTR"
                     /label="CMV-LTR"
     misc_feature    4560..5148
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /label="WPRE"
     misc_feature    1337..1570
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to
                     thecytoplasm."
                     /label="RRE"
     misc_feature    715..840
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /label="HIV-1 Psi"
     misc_feature    2062..2178
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     misc_feature    4413..4526
                     /label="3' flank"
     misc_feature    4206..4315
                     /label="5' flank"
     misc_feature    4316..4412
                     /label="IKZF1 sh1"
     misc_feature    3496..3525
                     /label="Gibson arm"
     misc_feature    4541..4570
                     /label="Gibson arm"
     misc_feature    4176..4205
                     /label="Gibson arm"
     misc_feature    4355..4376
                     /label="stem loop"
     misc_feature    3526..3545
                     /label="Primer anneal"
     misc_feature    complement(4169..4188)
                     /label="Primer anneal"
     misc_feature    4129..4148
                     /label="2186 PuroSeq"
     misc_feature    3458..3477
                     /label="Seq1"
     misc_feature    4189..>4205
                     /label="Gibson arm"
     misc_feature    4527..4540
                     /label="filler"
     misc_feature    2213..2214
                     /note="MCS"
                     /label="MCS"
     misc_feature    4189..4196
                     /note="Geneious type: restriction site"
                     /label="NotI"
     misc_feature    4546..4553
                     /note="Geneious type: restriction site"
                     /label="AscI"
     misc_feature    3723..3729
                     /note="Geneious type: restriction site"
                     /label="BstEII"
     misc_feature    4200..4205
                     /note="Geneious type: restriction site"
                     /label="BamHI"
     misc_feature    405..410
                     /note="Geneious type: restriction site"
                     /label="NheI"
     misc_feature    275..280
                     /note="Geneious type: restriction site"
                     /label="NdeI"
     misc_feature    2407..2412
                     /note="Geneious type: restriction site"
                     /label="NdeI"
     misc_feature    7299..7304
                     /note="Geneious type: restriction site"
                     /label="PvuI"
     misc_feature    7410..7415
                     /note="Geneious type: restriction site"
                     /label="ScaI"
     misc_feature    2213..2218
                     /note="Geneious type: restriction site"
                     /label="ClaI"
     misc_feature    5285..5290
                     /note="Geneious type: restriction site"
                     /label="KpnI"
     misc_feature    5815..5820
                     /note="Geneious type: restriction site"
                     /label="SphI"
     misc_feature    5743..5748
                     /note="Geneious type: restriction site"
                     /label="SphI"
     misc_feature    865..870
                     /note="Geneious type: restriction site"
                     /label="NruI"
     misc_feature    2512..2517
                     /note="Geneious type: restriction site"
                     /label="SnaBI"
     misc_feature    380..385
                     /note="Geneious type: restriction site"
                     /label="SnaBI"
     misc_feature    6020..6025
                     /note="Geneious type: restriction site"
                     /label="XmaI"
     misc_feature    6020..6025
                     /note="Geneious type: restriction site"
                     /label="SmaI"
     misc_feature    1223..1228
                     /note="Geneious type: restriction site"
                     /label="MfeI"
     misc_feature    5161..5166
                     /note="Geneious type: restriction site"
                     /label="PstI"
     misc_feature    467..472
                     /note="Geneious type: restriction site"
                     /label="PvuII"
     misc_feature    5314..5319
                     /note="Geneious type: restriction site"
                     /label="PvuII"
     misc_feature    2825..2830
                     /note="Geneious type: restriction site"
                     /label="AgeI"
     misc_feature    3526..3531
                     /note="Geneious type: restriction site"
                     /label="BamHI"
     misc_feature    1961..1966
                     /note="Geneious type: restriction site"
                     /label="AvaI"
     misc_feature    6020..6025
                     /note="Geneious type: restriction site"
                     /label="AvaI"
     misc_feature    3962..3967
                     /note="Geneious type: restriction site"
                     /label="BalI"
     misc_feature    3070..3075
                     /note="Geneious type: restriction site"
                     /label="SalI"
     misc_feature    7853..3
                     /note="Geneious type: restriction site"
                     /label="SalI"
     misc_feature    4284..4289
                     /note="Geneious type: restriction site"
                     /label="HpaI"
     misc_feature    4540..4545
                     /note="Geneious type: restriction site"
                     /label="MluI"
     misc_feature    4223..4228
                     /note="Geneious type: restriction site"
                     /label="ScaI"
     misc_feature    4301..4306
                     /note="Geneious type: restriction site"
                     /label="XhoI"
ORIGIN      
        1 gacggatcgg gagatcctcg cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accagctagc gaggcgtggc
      421 ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc
      481 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
      541 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
      601 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
      661 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
      721 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
      781 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
      841 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
      901 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
      961 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     1021 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     1081 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     1141 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     1201 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1261 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1321 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1381 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1441 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1501 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1561 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1621 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1681 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1741 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     1801 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1861 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     1921 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     1981 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2041 acggatctcg acggtatcgc ctttaaaaga aaagggggga ttggggggta cagtgcaggg
     2101 gaaagaatag tagacataat agcaacagac atacaaacta aagaactaca aaaacaaatt
     2161 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttatcgataa
     2221 gcttgggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2281 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2341 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2401 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     2461 cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta
     2521 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg
     2581 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2641 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2701 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     2761 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg actctagcta
     2821 gagtaccggt ccatggccca gtccaagcac ggcctgacca aggagatgac catgaagtac
     2881 cgcatggagg gctgcgtgga cggccacaag ttcgtgatca ccggcgaggg catcggctac
     2941 cccttcaagg gcaagcaggc catcaacctg tgcgtggtgg agggcggccc cttgcccttc
     3001 gccgaggaca tcttgtccgc cgccttcatg tacggcaacc gcgtgttcac cgagtacccc
     3061 caggacatcg tcgactactt caagaactcc tgccccgccg gctacacctg ggaccgctcc
     3121 ttcctgttcg aggacggcgc cgtgtgcatc tgcaacgccg acatcaccgt gagcgtggag
     3181 gagaactgca tgtaccacga gtccaagttc tacggcgtga acttccccgc cgacggcccc
     3241 gtgatgaaga agatgaccga caactgggag ccctcctgcg agaagatcat ccccgtgccc
     3301 aagcagggca tcttgaaggg cgacgtgagc atgtacctgc tgctgaagga cggtggccgc
     3361 ttgcgctgcc agttcgacac cgtgtacaag gccaagtccg tgccccgcaa gatgcccgac
     3421 tggcacttca tccagcacaa gctgacccgc gaggaccgca gcgacgccaa gaaccagaag
     3481 tggcacctga ccgagcacgc catcgcctcc ggctccgcct tgcccggatc cggcgcaaca
     3541 aacttctctc tgctgaaaca agccggagat gtcgaagaga atcctggacc gaccgagtac
     3601 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     3661 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     3721 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     3781 tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag cgtcgaagcg
     3841 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     3901 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     3961 ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     4021 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     4081 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     4141 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagc ggccgcaatg
     4201 gatcctgttt gaatgaggct tcagtacttt acagaatcgt tgcctgcaca tcttggaaac
     4261 acttgctggg attacttctt caggttaacc caacagaagg ctcgagaagg tatattgctg
     4321 ttgacagtga gcgcgctacg agaaggagaa cgaaatagtg aagccacaga tgtatttcgt
     4381 tctccttctc gtagcttgcc tactgcctcg gacttcaagg ggctacttta ggagcaatta
     4441 tcttgtttac taaaactgaa taccttgcta tctctttgat acatttttac aaagctgaat
     4501 taaaatggta taaattaaat cactttagct tgcagatcta cgcgtggcgc gcctggaaca
     4561 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4621 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4681 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4741 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4801 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4861 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4921 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     4981 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5041 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5101 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt
     5161 ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     5221 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttt ccagtcacac
     5281 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     5341 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     5401 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5461 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5521 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5581 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     5641 gaatatcaga gagtgagagg ccttgacatt gtttaaactg gaatgtgtgt cagttagggt
     5701 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5761 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5821 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5881 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5941 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6001 taggcttttg caaaaagctc ccgggagctt gtatatccat tacatgtgag caaaaggcca
     6061 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6121 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6181 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6241 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6301 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6361 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6421 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6481 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6541 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6601 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6781 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6841 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6901 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6961 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7021 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7081 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7141 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7201 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7261 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7321 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7381 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7441 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7501 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7561 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7621 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7681 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7741 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7801 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
